Establishment of new cell lines from Pseudaletia unipuncta with differential ...

1. An insect cell line established from embryonic egg cells from an insect from the order Lepidoptera which is designated BTI-Pu-527A7S and has been deposited under the accession number ATCC CRL 12285 and has the following characteristics:

a) supports replication of virus,

b) supports expression of protein after infection by a recombinant virus in a serum containing medium,

c) can grow in suspension and/or shaker flask cell cultures; and

d) grows in said serum containing medium and retains said ability to support replication of virus and to support expression of protein.

2. The insect cell line of claim 1 wherein at least 30 occlusion bodies are produced per cell line after infection by Autographa californica multiple nuclear polyhedrosis virus by the following steps:

a) seeding said cell line in at least one well at a density of 1.times.10.sup.5 cells/mL in TNM-FH medium;

b) allowing said cells to attach for two to three hours;

c) drawing off the old medium;

d) adding 0.5 mL of fresh TNM-FH medium containing said virus at a multiplicity of infection of five;

e) measuring said occlusion body production at six days post infection.

3. The insect cell line of claim 2 wherein at least 100 occlusion bodies are produced per cell line after infection by Autographa californica multiple nuclear polyhedrosis virus.

4. The insect cell line of claim 2 wherein at least 90 occlusion bodies are produced per cell line after infection by Autographa californica multiple nuclear polyhedrosis virus.

5. The insect cell line of claim 1 wherein at least 10,000 mean units (Unit/mL) of expressed Beta-galactosidase is produced after infection by a recombinant Autographa californica multiple nuclear polyhedrosis virus by the following steps:

a) seeding said cell line in at least one well at a density of 1.times.10.sup.5 cells/mL;

b) allowing said cells to attach for two to three hours;

c) drawing off the old medium;

d) adding 0.5 mL of fresh medium contain said recombinant virus at a multiplicity of infection of ten;

e) centrifuging said combination of cells, medium and virus at 1,000.times.g for 1 hour;

f) removing said media from said centrifuged combination; and

g) adding 0.5 mL of fresh medium;

h) measuring said Beta-galactosidase production at six days post infection.

6. An insect cell line with all of the identifying characteristics of the cell line identified as BTI-Pu-527A7S and deposited under the accession number ATCC CRL 12285.

7. An insect cell line established from the embryonic egg cells of the insect Pseudaletia unipuncta, from the order Lepidoptera, which is designated BTI-Pu-527A75 having all of the identifying characteristics of the insect cell line.